Journal: Applied Sciences

Article Title: Antiphotoaging Effect of (2′S)-Columbianetin from Corydalis heterocarpa in UVA-Irradiated Human Dermal Fibroblasts

doi: 10.3390/app10072568

Figure Lengend Snippet: Figure 5. Effect of (2′S)-columbianetin on the protein levels of; total and phosphorylated (p-) p38, ERK and JNK MAPKs (A), TGFβ, Smad2/3 (total and p-), Smad7 and Smad4 (B), and nuclear levels of p-Smad2/3 and Smad4 (C) in UVA irradiated HDFs following 24 h treatment. Protein levels were observed by Western blotting. Bands were quantified densitometrically and shown as relative percentage of UVA-irradiated untreated control group. β-actin and lamin B1 were used as internal loading control. Values are means ± SD (n = 3). # p < 0.05, ## p < 0.01 compared to non-irradiated blank group; * p < 0.05, ** p < 0.01 compared to UVA-irradiated untreated control group.

Article Snippet: Blocked membranes were washed with 1X TBST and incubated with primary antibodies (diluted 1:1000) against MMP-1 (sc-6837; Santa Cruz Biotechnology, Santa Cruz, CA, USA), type I pro-collagen (sc-8782; Santa Cruz Biotechnology), collagen (sc-29318; Santa Cruz Biotechnology), p38 (#8690; Cell Signaling Technology, Beverly, MA, USA), p-p38 (#4511; Cell Signaling Technology), JNK (LF-PA0047; Thermo Fisher Scientific), p-JNK (sc-293136; Santa Cruz Biotechnology), ERK (#4695; Cell Signaling Technology), p-ERK (#4370; Cell Signaling Technology), TGFβ (#3711; Cell Signaling Technology), Smad2/3 (sc-133098; Santa Cruz Biotechnology), p-Smad2/3 (sc-11769; Santa Cruz Biotechnology), Smad7 (sc-101152; Santa Cruz Biotechnology), Samd4 (sc-56479; Santa Cruz Biotechnology), lamin B1 (sc-374015; Santa Cruz Biotechnology) and β-actin (sc-47778; Santa Cruz Biotechnology) in primary antibody dilution buffer containing 1X TBST with 5% bovine serum albumin overnight at 4 ◦C.

Techniques: Irradiation, Western Blot, Control

Journal: FEBS letters

Article Title: MicroRNA-520g induces epithelial-mesenchymal transition and promotes metastasis of hepatocellular carcinoma by targeting SMAD7.

doi: 10.1016/j.febslet.2014.11.031

Figure Lengend Snippet: Fig. 4. SMAD7 is identified as a functional target of miR-520g in HCC. (A) qRT-PCR and (B) Western blot analysis of SMAD7 expression in HepG2 cells with miR-520g or control vectors transfection. n = 6; ⁄P < 0.05. (C) miR-520g and its putative binding sequence in the 30-UTR of SMAD7. The mutant miR-520g binding site was generated in the complementary site for the seed region of miR-520g (wt, wild type; mt, mutant type). (D) MiR-520g significantly suppressed the luciferase activity that carried wt but not mt 30-UTR of SMAD7. Anti-miR-520g led to a noticeable increase in luciferase activity of wt 30-UTR of SMAD7. n = 3 repeats with similar results; ⁄P < 0.05.

Article Snippet: Primer pairs against Homo sapiens gene SMAD7 (HP208950) and GAPDH (HP205798) were purchased from OriGene (Beijing, China).

Techniques: Functional Assay, Quantitative RT-PCR, Western Blot, Expressing, Control, Transfection, Binding Assay, Sequencing, Mutagenesis, Generated, Luciferase, Activity Assay

Journal: FEBS letters

Article Title: MicroRNA-520g induces epithelial-mesenchymal transition and promotes metastasis of hepatocellular carcinoma by targeting SMAD7.

doi: 10.1016/j.febslet.2014.11.031

Figure Lengend Snippet: Fig. 5. An inverse correlation between miR-520g and SMAD7 expression is observed in HCC. Comparing differences in the expression levels of SMAD7 (A) mRNA and (B) protein between HCC and matched non-tumor tissues. ⁄P < 0.05. (C) Representative immunostaining showed negative expression of SMAD7 in miR-520 high-expressing HCC tissue and positive expression of SMAD7 in miR-520 low-expressing tumor. A significant inverse correlation between miR-520g and SMAD7 expression was observed in HCC tissues. Scale bar: 100 lm; ⁄P < 0.05.

Article Snippet: Primer pairs against Homo sapiens gene SMAD7 (HP208950) and GAPDH (HP205798) were purchased from OriGene (Beijing, China).

Techniques: Expressing, Immunostaining

Journal: FEBS letters

Article Title: MicroRNA-520g induces epithelial-mesenchymal transition and promotes metastasis of hepatocellular carcinoma by targeting SMAD7.

doi: 10.1016/j.febslet.2014.11.031

Figure Lengend Snippet: Fig. 6. MiR-520g induces HCC cell migration, invasion and EMT by targeting SMAD7. (A) Western blot analysis of the expression of E-cadherin and vimentin in pCMV5- SMAD7 transfected miR-520g overexpressing HepG2 cells. n = 6; ⁄P < 0.05. (B) Migration and invasion abilities of the stable transfected cells following restoration of SMAD7 in miR-520g overexpressing HepG2 cells were analyzed by Boyden chamber and Transwell assays. n = 3 repeats with similar results; ⁄P < 0.05. (C) SMAD7 knockdown by a specific siRNA promoted the migration and invasion of miR-520g downregulating HCCLM3 cells. n = 3 repeats with similar results; ⁄P < 0.05.

Article Snippet: Primer pairs against Homo sapiens gene SMAD7 (HP208950) and GAPDH (HP205798) were purchased from OriGene (Beijing, China).

Techniques: Migration, Western Blot, Expressing, Transfection, Knockdown

Journal: Scientific Reports

Article Title: MiR-146a-5p suppresses activation and proliferation of hepatic stellate cells in nonalcoholic fibrosing steatohepatitis through directly targeting Wnt1 and Wnt5a

doi: 10.1038/srep16163

Figure Lengend Snippet: ( A ) Desmin staining indicating LX-2 and HSC-T6 cell proliferation decrease when miR-146a-5p was overexpressed. ( B ) miR-146a-5p inhibited LX-2 and HSC-T6 cell growth as determined by CCK-8 assays. Values are mean ± SD, * P < 0.05, ** P < 0.01 compared with mimics control. ( C ) Effect of miR-146a-5p on mRNA and ( D ) protein expression of fibrogenic genes α-SMA, Col-1, MMP-2 and Smad7. β-actin was used as loading control. Values are mean ± SD, ** P < 0.01 compared with control.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies against α-SMA, Col-1, MMP-2 (Bioss, Beijing, China), Smad7 (Novus Biologicals, Littleton, USA), Wnt1 (Abcam, Cambridge, MA, USA), Wnt5a (Novus Biologicals, Littleton, USA), β-catenin, GSK-3β (ProteinTech Group, Chicago, USA), NFAT5 (Santa Cruz, CA, USA).

Techniques: Staining, CCK-8 Assay, Control, Expressing

Journal: Scientific Reports

Article Title: MiR-146a-5p suppresses activation and proliferation of hepatic stellate cells in nonalcoholic fibrosing steatohepatitis through directly targeting Wnt1 and Wnt5a

doi: 10.1038/srep16163

Figure Lengend Snippet: ( A ) Knockdown efficiency of Wnt1 or Wnt5a in HSC-T6 cells was confirmed by RT-PCR and ( B ) Western blot. ( C ) Knockdown of Wnt1 or Wnt5a inhibits the mRNA and protein expressions ( D ) of β-catenin, NFAT5, and increase the expression of GSK-3β. ( E ) Knockdown of Wnt1 or Wnt5a inhibits the mRNA and protein levels (F) of α-SMA, Col-1 and MMP-2, and enhanced the expression of Smad7. β-actin was used as loading control. Values are mean ± SD, * P < 0.05, ** P < 0.01 compared with control.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies against α-SMA, Col-1, MMP-2 (Bioss, Beijing, China), Smad7 (Novus Biologicals, Littleton, USA), Wnt1 (Abcam, Cambridge, MA, USA), Wnt5a (Novus Biologicals, Littleton, USA), β-catenin, GSK-3β (ProteinTech Group, Chicago, USA), NFAT5 (Santa Cruz, CA, USA).

Techniques: Knockdown, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Control

Journal: Scientific Reports

Article Title: MiR-146a-5p suppresses activation and proliferation of hepatic stellate cells in nonalcoholic fibrosing steatohepatitis through directly targeting Wnt1 and Wnt5a

doi: 10.1038/srep16163

Figure Lengend Snippet: Primers for real-time quantitative PCR analysis.

Article Snippet: The membranes were incubated overnight at 4 °C with primary antibodies against α-SMA, Col-1, MMP-2 (Bioss, Beijing, China), Smad7 (Novus Biologicals, Littleton, USA), Wnt1 (Abcam, Cambridge, MA, USA), Wnt5a (Novus Biologicals, Littleton, USA), β-catenin, GSK-3β (ProteinTech Group, Chicago, USA), NFAT5 (Santa Cruz, CA, USA).

Techniques: Real-time Polymerase Chain Reaction